Exploring a reliable and sensitive method to detect telomere dysfunction in the aging population and premature-aging diseases

探索一种可靠、灵敏的方法来检测人口老龄化和早衰疾病中的端粒功能障碍

• 批准号: 10250901
• 负责人: Yie Liu
• 金额: $ 29.08万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10250901
• 关键词: Age; Aging; Aplastic Anemia; Biological Assay; Cell Aging; Cell Death; Chromosomes; DNA; DNA Damage; DNA strand break; Disease; Dyskeratosis Congenita; Elderly; Environmental Risk Factor; Fibroblasts; Fluorescent in Situ Hybridization; Functional disorder; Gamma-H2AX; Genetic Diseases; Genomic Instability; H2AFX gene; Human; Human Genetics; Human Herpesvirus 4; Immunofluorescence Immunologic; Link; Methods; Mus; Nucleoproteins; Organ; Pathogenesis; Premature aging syndrome; Primates; Proteins; Signal Transduction; Structure; Transformed Cell Line; age related; aging population; disorder risk; idiopathic pulmonary fibrosis; lifestyle factors; mortality; response; senescence; telomere; telomere loss

Telomere dysfunction-induced foci (TIF) are detected in senescent human fibroblasts and murine organs, and elderly primates and have recently been used as a marker for aging. Currently, TIF is determined by the colocalization of telomeric DNA or proteins with phosphorylated histone H2AX (gamma-H2AX) using immunofluorescence or immuno-telomere FISH analysis. However, due to the incapability of these conventional assays to detect TIFs at short telomeres, it is unclear if eroded telomeres in human conditions, such as aging and age-related diseases become dysfunctional, thereby contributing to DNA damage signaling and cellular senescence. We have developed a sensitive assay to accurately detect the colocalization of gamma-H2AX and telomeric DNA (or TIFs) in human fibroblast lines harboring critically short telomeres and telomeric DNA strand breaks. Currently, we are establishing the assay conditions in human EBV-transformed cell lines.

端粒功能障碍诱导的灶（TIF）在衰老的人成纤维细胞和鼠健身器中被检测到，以及老年灵长类动物，最近被用作衰老的标志。当前，TIF是通过使用免疫荧光或免疫 - 凝集素鱼类分析的端粒DNA或蛋白与磷酸化组蛋白H2AX（GAMMA-H2AX）共定位确定的。但是，由于这些常规测定能力在短端粒处检测TIF，因此尚不清楚在人类条件下侵蚀端粒是否侵蚀，例如衰老和与年龄相关的疾病的功能失调，从而有助于DNA损伤信号和细胞衰老。我们已经开发了一种敏感的测定，以准确检测到具有非常短的端粒和端粒DNA链断裂的人成纤维细胞中γ-H2AX和端粒DNA（或TIFS）的共定位。目前，我们正在建立人类EBV转换细胞系中的测定条件。